Study On The Function And Mechanism Of PIMREG In Glioma

Background: Glioma is the most common primary brain tumor in the human brain,accounting for about half of all intracranial primary tumors,with a high mortality rate.Although the comprehensive treatment of surgical resection,radiotherapy and chemotherapy has improved,the prognosis of patients with glioma is still poor.Therefore,it is urgent to find a new strategy for the treatment of glioma.PICALM interacting mitotic regulator（PIMREG）（NM₀19013）,also known as FAM64 A,CATS and RCS1,plays an important role in the regulation of cell proliferation,tumor invasion and metastasis.Studies have found that PIMREG can promote the proliferation and migration of breast cancer by enhancing NF-κB signal activation,and inhibiting PIMREG can inhibit the growth of breast cancer cells.Up-regulation of PIMREG can also promote the growth and metastasis of osteosarcoma cells.The GEPIA database shows that PIMREG is significantly up-regulated in glioma patients.Therefore,we speculate that PIMREG plays an important role in the occurrence and development of glioma.In summary,this project intends to study the role of PIMREG in glioma and its potential molecular mechanism.Objective: To study the expression and biological functions of PIMREG in different glioma cells,as well as the molecular mechanism of PIMREG on β-catenin signaling pathway regulating the proliferation,migration and invasion of glioma cells,and to explore the role of PIMREG in glioma The mechanism of action in PIMREG provides a theoretical basis for PIMREG as a new therapeutic target for glioma.Methods:（1）By using the TCGA database to analyze the expression of PIMREG in human glioma,Real-time PCR and Western Blot verify the expression of PIMREG in the human glioma cell lines U251,T98 G,SHG-44,and A172.（2）Transfect the PIMREG overexpression plasmid into a low-expressing glioma cell line,and simultaneously transfect two PIMREG interference fragments into a high-expressing glioma cell line,and use Real-time PCR and Western Blot to detect PIMREG and cyclin The expression of D1 and cyclin E,CCK-8 to detect cell proliferation and flow cytometry to detect cell cycle distribution.（3）Detect the effect of PIMREG on the migration and invasion ability of glioma cells by scratch test and transwell invasion test.At the same time,Western blot was used to detect the expression of mature MMP-2,mature MMP-9,E-cadherin and Vimentin in the cells.（4）Western blot was used to detect the expression of β-catenin and c-myc in over-expressed and under-expressed PIMREG glioma cells,while immunofluorescence staining was used to detect the expression of β-catenin in over-expressed and under-expressed PIMREG glioma cells Express the situation.（5）After the β-catenin inhibitor（ICG-001）was treated with PIMREG overexpressing glioma cells for 24 hours,the cell proliferation and invasion ability were detected by CCK-8 and transwell invasion experiments,and Western blot was used to detect MMP-2.The expression of Vimentin and cyclin D1.Results:（1）By using the TCGA data set analysis tool "Gene Expression Profile Interactive Analysis"（GEPIA2）,we found that PIMREG is highly expressed in human brain glioma tissue.And by Western blot detection of different human glioma cell lines U251,T98 G,SHG-44 and A172,the expression of PIMREG increased in sequence.（2）CCK-8 experiment results show that overexpression of PIMREG can significantly promote the proliferation of glioma cells,and inhibition of PIMREG expression can significantly inhibit the proliferation of glioma cells.The cell cycle distribution was analyzed by flow cytometry.The overexpression of PIMREG significantly promoted the proliferation of glioma cells and promoted the transition from G1 phase to S phase.Western blot detection showed that after PIMREG was overexpressed,cyclin D1 and cyclin The expression of E is up-regulated.After suppressing the expression of PIMREG,glioma cells showed the opposite effect.（3）Through the cell scratch test and transwell invasion test,the up-regulation of PIMREG significantly enhanced the migration and invasion ability of glioma cells,and the results of Western blot showed that the up-regulation of PIMREG reduced the expression of E-cadherin and increased the expression of Vimentin.MMP-2 and MMP-9 expression.After suppressing the expression of PIMREG,glioma cells showed the opposite effect.（4）Through Western blot experiment,the expression of β-catenin and c-myc was significantly enhanced after PIMREG overexpression.The results of immunofluorescence experiments were obvious,and the expression of β-catenin increased significantly after PIMEREG was overexpressed.After suppressing the expression of PIMREG,glioma cells showed the opposite effect.（5）After treating glioma cells with β-catenin inhibitor ICG-001 for 24 hours,CCK-8 cell proliferation experiments and transwell invasion experiments showed that the proliferation and invasion ability of glioma cells decreased,and the results of Western blot experiments showed that Vimentin,The expression of MMP-2 and cyclin D1 decreased.Conclusion:This study revealed that PIMREG can promote the proliferation,migration and invasion of glioma cells by regulating the β-catenin signaling pathway.This research may help to explore the pathogenesis of glioma and new targets for glioma treatment.